Abstract: Random Amplified Polymorphic DNA(RAPD) markers were applied to analyze 23 Osmanthus fragrans cultivars in Guilin City. Among the total 193 fragments generated by 20 selected primers (among 100 primers) , 114 appeared to be polymorphic(59. 1%). Cluster analysis based on the RAPD results was performed and dendrogram was constructed. These cultivars tested by RAPD were divided into 4 cultivar groups. The result was consistent with that from traditional taxonomy analysis. The RAPD study illustrated that it is possible to analyze intra-cultivar variation of O. fragrans cultivars on the basis of genotype rather than phenotype and therefore, offered significant evidences in solving taxonomic problem of O. fragrans cultivars in Guilin City.